文心兰OnGI在拟南芥中异源表达促进开花

doi:10.16420/j.issn.0513-353x.2021-0182

摘要/Abstract

摘要：

对文心兰（Oncidium）进行不同的光周期处理,结果发现长日照条件有利于花芽的形成;克隆获得文心兰OnGI基因,其编码区长3 483 bp,编码1 161个氨基酸;多序列比对结果显示其与石斛（Dendrobium nobile）DnGI同源性最高,为86.57%;进化树分析结果表明其与其他兰科植物为同一组。组织表达分析结果显示,OnGI在文心兰花中表达量最高,尤其在花苞形成初始时期;在不同生长时期的叶片中,花芽分化期OnGI表达量达到峰值;在不同光周期条件下,OnGI的表达模式存在差异,与拟南芥（Arabidopsis thaliana）类似。亚细胞定位显示,OnGI定位在细胞核中。在拟南芥中过表达OnGI可以提早开花。根据结果推测,文心兰和拟南芥可能存在类似的光周期调控GI表达的机制,调控植株开花。

关键词: 文心兰, OnGI, 光周期, 表达, 开花

Abstract:

In this study,it was found that long-day condition could promote the flower bud differentiation of Oncidium under different photoperiod treatments. The OnGI gene of Oncidium was cloned,and its coding region was 3 483 bp long,encoding 1 161 amino acids. The results of multiple sequence alignment showed that it had the highest homology with Dendrobium nobile DnGI（86.57%）. The phylogenetic tree analysis showed that it was classified into the same group with other orchids. Tissue expression analysis showed that the expression of OnGI was the highest in flowers,especially at the initial stage of bud formation,and in leaves at different growth stages,the expression of OnGI peaked at the stage of flower bud differentiation. Under different photoperiod conditions,the expression pattern of OnGI was different,similar to that of Arabidopsis thaliana. The results of subcellular localization showed that OnGI was localized in the nucleus. Overexpression of OnGI in Arabidopsis thaliana could lead to earlier flowering. Based on the above results,we speculate that there may be a similar photoperiod regulation mechanism of GI expression in Arabidopsis thaliana and to regulate plant flowering.

Key words: Oncidium, OnGI, photoperiod, expression, flowering

中图分类号:

S682.31

方能炎, 樊荣辉, 罗远华, 孔兰, 林榕燕, 叶秀仙, 林兵, 钟淮钦, 黄敏玲. 文心兰OnGI在拟南芥中异源表达促进开花[J]. 园艺学报, 2022, 49(4): 841-850.

FANG Nengyan, FAN Ronghui, LUO Yuanhua, KONG Lan, LIN Rongyan, YE Xiuxian, LIN Bing, ZHONG Huaiqin, HUANG Minling. The Heterologous Expression of Oncidium OnGI Promotes Flowering in Arabidopsis thaliana[J]. Acta Horticulturae Sinica, 2022, 49(4): 841-850.

图/表 8

表1 引物序列

Table 1 Sequences of primers

用途 Application	引物名称 Primer name	引物序列（5′-3′） Sequence
克隆 Clone	OnGI-F	ATGTCTGCATCGTGTCAGAAGTG
	OnGI-R	CTAGCTGGGAATACGAAAACCCAA
转基因植株验证 Validation of transgenic plants	pEarleyGate104-Ver-F	CAGATCTCGAGCTCAAGCTTCGAAT
转基因植株验证 Validation of transgenic plants	pEarleyGate104-Ver-R	ACAACGTGCACAACAGAATTGAAAGC
qPCR	OnGI1-qF	GCAAGAGATGATGAGCAAGG
	OnGI1-qR	TAAGTGTCTCAGGGCTTCC
	OnActin-qF	AATGTGCCTGCTATGTATGTTGCT
	OnActin-qR	GGGCATATCCTTCGTAGATTGT
半定量PCR	RT-OnGI-F	AGCAAGGTCAAAGAATGTGGAGGAT
Semi-quantitative RT-PCR	RT-OnGI-R	TGTCTGAATGCCAGTTGGTGATGTA
	RT-AtGI-F	CTGTCTTTCTCCGTTGTTTCACTGT
	RT-AtGI-R	TCATTCCGTTCTTCTCTGTTGTTGG
	RT-AtTUB-F	CTCAAGAGGTTCTCAGCAGTA
	RT-AtTUB-R	TCACCTTCTTCATCCGCAGTT

图1 光周期对文心兰花芽分化率的影响

Fig. 1 Effect of photoperiod on bud formation rate of Oncidium

图2 OnGI扩增产物琼脂糖电泳图

Fig. 2 Agarose electrophoresis of OnGI amplification products

图3 不同植物中GI蛋白进化树分析百分数代表蛋白位点覆盖率。

Fig. 3 Phylogenetic analysis of the GI proteins from different plant species Percentage represents protein site coverage.

图4 文心兰中OnGI的表达 A. 不同组织;B. 不同生长时期的叶片;C. 不同发育时期的花。不同小写字母代表差异显著（P < 0.05）。

Fig. 4 Expression analysis of OnGI A. Different tissues;B. In leaves at different developmental stages;C. In flowers at different developmental stages. Different lowercase letters represent significant difference（P < 0.05）.

图5 不同光周期处理下OnGI 在叶片中表达分析

Fig. 5 Expression analysis of OnGI in leaves under different photoperiods

图6 OnGI亚细胞定位

Fig. 6 Subcellular localization of OnGI

图7 OnGI在转基因拟南芥植株中的表达（A）及表型（B、C） WT:野生型;OE-1 ~ OE-3:过表达转基因株系。

Fig. 7 The expression of OnGI in transgenic Arabidopsis thaliana plants（A）and the phenotype（B,C） WT:Wild type;OE-1-OE-3:OnGI overexpression of transgenic Arabidopsis thaliana lines.

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